Abstract
Eukaryotic translation initiation factor eIF-4A is a member of the DEAD box family of RNA helicases and RNA-dependent ATPases. In tobacco, eIF-4A is encoded by a gene family with one isoform, eIF-4A8, being exclusively expressed in pollen. This pollen-specific isoform is a candidate for mediating translational control in the developing gametophyte. Here we show that eIF-4A is barely phosphorylated in mature pollen, but during pollen tube germination, two isoforms of eIF-4A become phosphorylated. Phosphoamino acid analysis indicated phosphorylation of threonine. In order to determine whether pollen-specific eIF-4A8 is among the phosphorylated isoforms, we raised transgenic tobacco plants overexpressing eIF-4A8 containing a histidine tag. Hereby, we could show that indeed eIF-4A8 is modified through phosphorylation. The biological relevance of the phosphorylation of eIF-4A is discussed.
Full Text
The Full Text of this article is available as a PDF (161.6 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bate N., Spurr C., Foster G. D., Twell D. Maturation-specific translational enhancement mediated by the 5'-UTR of a late pollen transcript. Plant J. 1996 Oct;10(4):613–623. doi: 10.1046/j.1365-313x.1996.10040613.x. [DOI] [PubMed] [Google Scholar]
- Boyle W. J., van der Geer P., Hunter T. Phosphopeptide mapping and phosphoamino acid analysis by two-dimensional separation on thin-layer cellulose plates. Methods Enzymol. 1991;201:110–149. doi: 10.1016/0076-6879(91)01013-r. [DOI] [PubMed] [Google Scholar]
- Brander K. A., Kuhlemeier C. A pollen-specific DEAD-box protein related to translation initiation factor eIF-4A from tobacco. Plant Mol Biol. 1995 Feb;27(4):637–649. doi: 10.1007/BF00020219. [DOI] [PubMed] [Google Scholar]
- Brander K. A., Mandel T., Owttrim G. W., Kuhlemeier C. Highly conserved genes coding for eukaryotic translation initiation factor eIF-4A of tobacco have specific alterations in functional motifs. Biochim Biophys Acta. 1995 Apr 26;1261(3):442–444. doi: 10.1016/0167-4781(95)00052-i. [DOI] [PubMed] [Google Scholar]
- Depicker A., Stachel S., Dhaese P., Zambryski P., Goodman H. M. Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet. 1982;1(6):561–573. [PubMed] [Google Scholar]
- Duncan R. F., Hershey J. W. Initiation factor protein modifications and inhibition of protein synthesis. Mol Cell Biol. 1987 Mar;7(3):1293–1295. doi: 10.1128/mcb.7.3.1293. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Duncan R. F., Hershey J. W. Translational repression by chemical inducers of the stress response occurs by different pathways. Arch Biochem Biophys. 1987 Aug 1;256(2):651–661. doi: 10.1016/0003-9861(87)90622-9. [DOI] [PubMed] [Google Scholar]
- Gallie D. R., Le H., Caldwell C., Tanguay R. L., Hoang N. X., Browning K. S. The phosphorylation state of translation initiation factors is regulated developmentally and following heat shock in wheat. J Biol Chem. 1997 Jan 10;272(2):1046–1053. doi: 10.1074/jbc.272.2.1046. [DOI] [PubMed] [Google Scholar]
- Goldberg R. B., Beals T. P., Sanders P. M. Anther development: basic principles and practical applications. Plant Cell. 1993 Oct;5(10):1217–1229. doi: 10.1105/tpc.5.10.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hershey J. W. Protein phosphorylation controls translation rates. J Biol Chem. 1989 Dec 15;264(35):20823–20826. [PubMed] [Google Scholar]
- Langland J. O., Langland L. A., Browning K. S., Roth D. A. Phosphorylation of plant eukaryotic initiation factor-2 by the plant-encoded double-stranded RNA-dependent protein kinase, pPKR, and inhibition of protein synthesis in vitro. J Biol Chem. 1996 Feb 23;271(8):4539–4544. doi: 10.1074/jbc.271.8.4539. [DOI] [PubMed] [Google Scholar]
- Linder P., Slonimski P. P. An essential yeast protein, encoded by duplicated genes TIF1 and TIF2 and homologous to the mammalian translation initiation factor eIF-4A, can suppress a mitochondrial missense mutation. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2286–2290. doi: 10.1073/pnas.86.7.2286. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mandel T., Fleming A. J., Krähenbühl R., Kuhlemeier C. Definition of constitutive gene expression in plants: the translation initiation factor 4A gene as a model. Plant Mol Biol. 1995 Dec;29(5):995–1004. doi: 10.1007/BF00014972. [DOI] [PubMed] [Google Scholar]
- McCormick S. Male Gametophyte Development. Plant Cell. 1993 Oct;5(10):1265–1275. doi: 10.1105/tpc.5.10.1265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merrick W. C. Mechanism and regulation of eukaryotic protein synthesis. Microbiol Rev. 1992 Jun;56(2):291–315. doi: 10.1128/mr.56.2.291-315.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Minich W. B., Balasta M. L., Goss D. J., Rhoads R. E. Chromatographic resolution of in vivo phosphorylated and nonphosphorylated eukaryotic translation initiation factor eIF-4E: increased cap affinity of the phosphorylated form. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7668–7672. doi: 10.1073/pnas.91.16.7668. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morley S. J., Dever T. E., Etchison D., Traugh J. A. Phosphorylation of eIF-4F by protein kinase C or multipotential S6 kinase stimulates protein synthesis at initiation. J Biol Chem. 1991 Mar 15;266(8):4669–4672. [PubMed] [Google Scholar]
- Nielsen P. J., Trachsel H. The mouse protein synthesis initiation factor 4A gene family includes two related functional genes which are differentially expressed. EMBO J. 1988 Jul;7(7):2097–2105. doi: 10.1002/j.1460-2075.1988.tb03049.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Owttrim G. W., Hofmann S., Kuhlemeier C. Divergent genes for translation initiation factor eIF-4A are coordinately expressed in tobacco. Nucleic Acids Res. 1991 Oct 25;19(20):5491–5496. doi: 10.1093/nar/19.20.5491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Owttrim G. W., Mandel T., Trachsel H., Thomas A. A., Kuhlemeier C. Characterization of the tobacco eIF-4A gene family. Plant Mol Biol. 1994 Dec;26(6):1747–1757. doi: 10.1007/BF00019489. [DOI] [PubMed] [Google Scholar]
- Pain V. M. Initiation of protein synthesis in eukaryotic cells. Eur J Biochem. 1996 Mar 15;236(3):747–771. doi: 10.1111/j.1432-1033.1996.00747.x. [DOI] [PubMed] [Google Scholar]
- Pause A., Belsham G. J., Gingras A. C., Donzé O., Lin T. A., Lawrence J. C., Jr, Sonenberg N. Insulin-dependent stimulation of protein synthesis by phosphorylation of a regulator of 5'-cap function. Nature. 1994 Oct 27;371(6500):762–767. doi: 10.1038/371762a0. [DOI] [PubMed] [Google Scholar]
- Taylor Loverine P., Hepler Peter K. POLLEN GERMINATION AND TUBE GROWTH. Annu Rev Plant Physiol Plant Mol Biol. 1997 Jun;48(NaN):461–491. doi: 10.1146/annurev.arplant.48.1.461. [DOI] [PubMed] [Google Scholar]
- Warmke H. E., Lee S. L. Pollen Abortion in T Cytoplasmic Male-Sterile Corn (Zea mays): A Suggested Mechanism. Science. 1978 May 5;200(4341):561–563. doi: 10.1126/science.200.4341.561. [DOI] [PubMed] [Google Scholar]
- Webster C., Gaut R. L., Browning K. S., Ravel J. M., Roberts J. K. Hypoxia enhances phosphorylation of eukaryotic initiation factor 4A in maize root tips. J Biol Chem. 1991 Dec 5;266(34):23341–23346. [PubMed] [Google Scholar]
- Weinstein D. C., Honoré E., Hemmati-Brivanlou A. Epidermal induction and inhibition of neural fate by translation initiation factor 4AIII. Development. 1997 Nov;124(21):4235–4242. doi: 10.1242/dev.124.21.4235. [DOI] [PubMed] [Google Scholar]
